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Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization

Hernandez, Maria
Recalde, Sergio
Garcia-Garcia, Laura
Bezunartea, Jaione
Miskey, Csaba
Johnen, Sandra
Sebe, Attila
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Published in Molecular Therapy - Methods & Clinical Development. 2019, vol. 15, p. 403-417
Abstract Age-related macular degeneration (AMD) is a progressive retinal disorder characterized by imbalanced pro- and antiangiogenic signals. The aim of this study was to evaluate the effect of ex vivo cell-based gene therapy with stable expression of human pigment epithelium-derived factor (PEDF) release using the non-viral Sleeping Beauty (SB100X) transposon system delivered by miniplasmids free of antibiotic resistance markers (pFAR4). Retinal pigment epithelial (RPE) cells and iris pigment epithelial (IPE) cells were co-transfected with pFAR4-inverted terminal repeats (ITRs) CMV-PEDF-BGH and pFAR4-CMV-SB100X-SV40 plasmids. Laser-induced choroidal neovascularization (CNV) was performed in rats, and transfected primary cells (transfected RPE [tRPE] and transfected IPE [tIPE] cells) were injected into the subretinal space. The leakage and CNV areas, vascular endothelial growth factor (VEGF), PEDF protein expression, metalloproteinases 2 and 9 (MMP-2/9), and microglial/macrophage markers were measured. Injection with tRPE/IPE cells significantly reduced the leakage area at 7 and 14 days and the CNV area at 7 days. There was a significant increase in PEDF and the PEDF/VEGF ratio with tRPE cells and a reduction in the MMP-2 activity. Our data demonstrated that ex vivo non-viral gene therapy reduces CNV and could be an effective and safe therapeutic option for angiogenic retinal diseases.
Keywords Epithelium-derived factorRetinal-pigment epitheliumEndothelial growth-factorMacular degenerationRetroviral integrationTissue inhibitorSite selectionRCS ratsIrisPEDFAngiogenesisIris pigment epithelial cells (IPE)Retinal pigment epithelium (RPE)Pigment epithelium derived factor (PEDF)Vascular endothelial growth factor (VEGF)SB100X transposase
PMID: 31890733
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Research group Ophtalmologie expérimentale (925)
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HERNANDEZ, Maria et al. Preclinical Evaluation of a Cell-Based Gene Therapy Using the Sleeping Beauty Transposon System in Choroidal Neovascularization. In: Molecular Therapy - Methods & Clinical Development, 2019, vol. 15, p. 403-417. doi: 10.1016/j.omtm.2019.10.013 https://archive-ouverte.unige.ch/unige:138789

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Deposited on : 2020-07-31

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